Sulforhodamine B Assay: A Sensitive Assay to Measure Drug Resistance Via Determination of Cellular Protein Content in Cancer Cells

Abstract

Source: Sciarrillo, R. et al. Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models. J. Vis. Exp. (2016)

This video describes the high throughput colorimetric screening assay, sulforhodamine B assay, to measure drug-induced cytotoxicity in cancer cells. The assay determines the drug resistance by measuring the cell density based on the cellular protein content.

Protocol

1. SRB Assay for Pancreatic Carcinoma Cells Dissolve SRB reagent in 1% acetic acid at final concentration of 0.4% (w/v). Dissolve trichloroacetic acid (TCA) in ultrapure water at a final concentration of 50% (w/v). Dissolve Tris(hydroxymethyl)-aminomethane in ultrapure water at a final concentration of 10 mM. Prepare a separate 96-well flat-bottom "Day 0" control plate to ensure more accurate estimation of growth inhibition: Seed 6 wells with cells growin…

Materials

Sulforhodamine B	Sigma-Aldrich	230162
Trichloroacetic acid	Sigma-Aldrich	251399
Panc-1	ATCC, Manassas, VA, USA	ATCC CRL-1469
Tris(hydroxymethyl)-aminomethane	Sigma Aldrich	252859
Greiner CELLSTAR 96 well plates	Greiner/Sigma	M0812-100EA
Anthos-Elisa-reader 2001	Labtec, Heerhugowaard, Netherlands		UV-Vis 96-well plate spectrophotometer
RPMI-1640	Gibco, Carlsbad, CA, USA	11875093
Greiner CELLSTAR 96 well plates	Greiner/Sigma	M0812-100EA
Fetal bovine and calf serum	Greiner Bio-One, Frickenhausen, Germany	758093
penicillin G streptomycin sulphate	Gibco, Carlsbad, CA, USA	15140122